3011 HA Tag

Gene synthesis

Mix 1 ul of 100 uM H1 ~ H14, take 1 ul mix dilute to 10 ul with H2O

Mix 1 ul of 100 uM L1 ~L12, take 1 ul mix dilute to 10 ul with H2O

PCR

1 ul diluted mix,

5 ul 5X Q5 buffer

1 ul 10 mM dNTP

1 ul home-made-phusion

1 ul 10 uM H1, 1 ul  10 uM H14 for VH

or 1 ul 10 uM L1, 1 ul 10 uM L12 for VL

H2O to 25 ul

PCR

94 oC 2 min

25 cycles of 94 oC 30, 55 oC 30, 72 oC 45 .

72 oC 3 min.

9/22/2017

Cut out band, purify ligate to pJet1.2

miniprep, digest with Xba1 + Xho1

New 3011 synthesized by Eurofins--Paul.xu (talk) 10:02, 6 November 2017 (EST)--Paul.xu (talk) 10:02, 6 November 2017 (EST), 11/6/2017